• Proceedings of the Ginseng society Conference
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• 1974.09a
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• pp.9-22
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• 1974

Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

• Weed & Turfgrass Science
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• v.4 no.4
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• pp.348-359
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• 2015

Total 443 isolates of actinomycetes were isolated from turfgrass rhizosphere as potential biological control agents. The two isolates (S11 and S4) showed highest cellulase activity with compared to the other isolates that exhibited a clear zone of 1.2 mm around the colony on cellulose agar medium. S12 strain appeared the most active chitin degrading, which exhibited a 1.2 mm of clear zone. The highest proteolytic activity on skim milk agar was which exhibited a 7.5 mm of clear zone by S2 strain. S1 strain from the soli showed siderophore production ability, which exhibited a 0.6 mm of large clear zone on chrome azurol S agar. The antifungal activity of the volatile compound producing by 4 selected actinomycetes was investigated that inhibition rate against Rhizoctonia solani AG2-2 and Sclerotinia homoeocarpa. Growth inhibition effect of S8 isolate against S. homoeocarpa was appeared to 94.8%, S2 to 76.9%, S5 to 46.1% and S12 to 43.5%. The significant inhibition effects on mycelial growth of S. homoeocarpa were shown on media with four strains. The inhibition effect was the highest with S8 strain treatment at 94.8%.

• Journal of Life Science
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• v.21 no.4
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• pp.554-560
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• 2011

This study was performed to investigate the effects of microbial augmentation on the biological treatment of paper mill wastewater. Three bacteria (KN11, KN13, KN27) capable of degrading aromatic compounds and a bacterial strain (GT21) producing an extracellular cellulase were isolated from soil and wastewater by selective enrichment culture. Through morphological, physiological, and biochemical taxonomies, isolated strains of KN11, KN13, KN27, and GT21 were identified as Acinetobacter sp., Neisseria sp., Bacillus sp., and Pseudomonas sp. and named Acinetobacter sp. KN11, Neisseria sp. KN13, Bacillus sp. KN27, and Pseudomonas sp. GT21, respectively. For analysis of non-biodegradable and chemical oxygen demand (COD)-increasing matter in a paper mill wastewater, we utilized GC/MS to detect aromatic compounds and their derivatives containing several substituted functional groups. The microbial augmentation, J30 formulated with the mixture of bacteria including Acinetobacter sp. KN11, Neisseria sp. KN13, Bacillus sp. KN27, and Pseudomonas sp. GT21, was used for the treatment of paper mill wastewater. The optimum temperature and pH for COD removal of the microbial augmentation, J30, were $30^{\circ}C$ and 7.5, respectively. For evaluation of the industrial applicability of the microbial augmentation, J30 in the pilot test, treatment efficiency was examined using paper mill wastewater. The microbial augmentation, J30, showed a COD removal rate of 87%. On the basis of the above results, we designed the wastewater treatment process of the activated sludge system.

• Korean Journal of Food Science and Technology
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• v.29 no.5
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• pp.1028-1032
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• 1997

The strain No. 33, which produces cellulose-degrading enzyme, was isolated from soil. Yellow halo was identified when the culture supernatant of the strain was loaded onto agar plate containing 2.0% CMC using paper disc method. From scanning electron microscopic observation, the morphology of the stain was rod-shaped. For identification, several biochemical characteristics were tested, and this strain was identified to Bacillus sp. So, we named this strain as Bacillus sp. No. 33. The maximal growth was observed when the stain was cultured in the medium containing 1.0% glucose, 3.0% yeast extract, 0.5% $KH_2PO_4$, 0.02% $MgSO_4{\cdot}7H_2O$, pH 7.0 at $30^{\circ}C$ for 39 hours with shaking. The maximal enzyme production was accomplished using the medium containing 4.0% CMC, 2.0% yeast extract, 0.5% $KH_2PO_4$, 0.04% $MgSO_4{\cdot}7H_2O$, pH 7.0 at $30^{\circ}C$ for 42 hours with shaking.

• Korean Journal of Microbiology
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• v.31 no.1
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• pp.1-8
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• 1993

lntergeneric hybrids formed between Aspergillus niger and Penicillium verruculosum were obtained by nuclear transfer technique. Nuclei isolated from wild type and auxotrophic mutants of donor strains were transferred into the protoplasts of different auxotrophic mutants as recipient strains. Several auxotrophic mutants were isolated from conidiospores of the two strains mutagenized with ultraviolet and N-methyl-N'-nitrosoguanidine. Optimal conditions for formation of intergeneric hybrids were investigated. Frequencies of intergeneric hybrid formation by nuclear transfer were $7{\times}10^{5}~1{\times}10^{5}$. From observations of genetic stability. DNA content. nuclear stain and conidial size. it was suggested that their karyotypes are aneuploid. In addition. the hybrids possess the 1.1~2.3-fold higher cellulase activities than those of parental strains. It was also revealed that some hybrids had different isozyme patterns compared to those of parental strains by CMCase and $\beta$-glucosidase activity assays.

• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.119-125
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• 1988

$\beta$-Glucosidase of Cellulomonas sp. CS1-1 in cellular compartment was localized with cell-bound form while Avicelase and carboxymethylcellulase (CMCase) were appeared with extracellular enzyme. Cell growth on cellulose or CMC minimal broth was increased by glucose addition. $\beta$-Glucosidase production on cellobiose or CMC minimal broth was repressed by the addition of glucose. However, on CMC minimal broth, the enzyme production was specially stimulated by cellobiose addition. $\beta$-Glucosidase production was also induced by CMC, starcth and maltose compared with glycerol, arabinose, xylose and trehalose. From the above results, it was concluded that glucose effect on $\beta$-glucosidase biosynthesis showed catabolite repression, but enzyme production was induced by cellobiose, CMC, and starch, indicating that $\beta$-glucosidase is inducible enzyme. Yeast extract stimulated $\beta$-glucosidase production more than peptone and ammonium sulfate. $\beta$-Glucosidase activity was increased with 50mM MgCl$_2$in 10mM potassium phosphate buffer (pH 7.0). Optimum conditions for enzyme activities were pH 6.0 and 42$^{\circ}C$, Km value of $\beta$-glucosidase for p-nitrophenyl-$\beta$-D-glucosidase was 0.256mM and Ki for $\beta$-D(+)-glucose was 9.0mM.

• Journal of Korean Society of Forest Science
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• v.98 no.3
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• pp.305-310
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• 2009

Yellow poplar was selected a promising biomass resources for bio-ethanol production through alkali prehydrolysis, enzymatic saccharification and fermentation using commercial cellulase mixtures (Celluclast 1.5L and Novozym 342 mixtures) and fermenting yeast. In alkali prehydrolysis, 51.1% of Yellow poplar biomass remained as residues, which chemical compositions were 82.2% of cellulose, 17.6% of xylan and 2.0% of lignin. In alkali prehydrolysis process, 96.9% of cellulose, 38.0% of xylan and 5.7% of lignin were remained. Enzymatic saccharification by commercial cellulases led to 87.0% of cellulose to glucose and 87.2% of xylan to xylose conversion. Produced glucose and xylose were fermented with fermenting yeast (Saccharomycess cerevisiae), which resulted in selective fermentation of glucose only to bio-ethanol. Residual monosaccharides after fermentation were consisted to 0.4-1.4% of glucose and 92.1-99.5% of xylose. Ethanol concentration was highest for 24 h fermentation as 57.2 g/L, but gradually decreased to 56.2 g/L for 48 h fermentation and 54.3 g/L for 72 h fermentation, due to the ethanol consumption by fermenting yeast.

• Journal of Life Science
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• v.17 no.11
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• pp.1555-1561
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• 2007

In order to know the effects of scoria, germanium, charcoal, ginger, stevia, and CLA(Conjugated Linoleic Acid) as biologically active materials on pathogenic microbes and rumen anaerobic microbes, the growth rate of pathogens (including Escherichia coli O157, Salmonella paratyphi, Listeria monocytogenes and Staphylococcus aureus) and in vitro lumen microbial growth, gas production, ammonia concentration, carboxymethyl-cellulase (CMCase) activity, and microbial populations were investigated. The growth of pathogenic microbes was inhibited by the supplement of 0.10% ginger. Ginger had powerful antimicrobial properties on all the pathogens used in this experiments. Additionally in the antibacterial assay by paper disc method, we could observe the clear zone of similar area with the positive control(antibiotics) for E. coli as applied with the 10% stevia or the 10% CLA only. The supplements of ginger, stevia and CLA in vitro rumen fermentation inhibited populations of rumen bacteria and protozoa. Particularly supplement of ginger resulted in remarkable reduction of the protozoa population, which means it might serve as a source inhibiting material of methane creation in the rumen.

• Microbiology and Biotechnology Letters
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• v.40 no.1
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• pp.30-38
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• 2012

We investigated the effects on soil microbial diversity and the growth promotion of red pepper resulting from inoculation with a microbial agent composed of Bacillus subtilis AH18, B. licheniformis K11 and Pseudomonas fluorescens 2112 in a red pepper farming field. Photosynthetic bacteria, Trichoderma spp., Azotobacter spp., Actinomycetes, nitrate oxidizing bacteria, nitrite oxidizing bacteria, nitrogen fixing bacteria, denitrifying bacteria, phosphate solubilizing bacteria, cellulase producing bacteria, and urease producing bacteria are all indicator microbes of healthy soil microbial diversity. The microbial diversity of the consortium microbial agent treated soil was seen to be 1.1 to 14 times greater than soils where other commercial agent treatments were used, the latter being the commercial agent AC-1, and chemical fertilizer. The yield of red pepper in the field with the treated consortium microbial agent was increased by more than 15% when compared to the other treatments. Overall, the microbial diversity of the red pepper farming field soil was improved by the consortium microbial agent, and the promotion of growth and subsequent yield of red pepper was higher than soils where the other treatments were utilized.

• Applied Biological Chemistry
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• v.37 no.5
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• pp.343-348
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• 1994

Ti plasmid of A. tumefaciens was labeled with $^3H-thymidine$, purified and encapsulated into phosphatidylserine (PS) and PS-cholesterol (Chol; 1 : 1 molar ratio) liposomes by lyophilization-rehydration method. PS was supplemented with 1 mole percent octadecyl rhodamine B for fluorometric measurement of PS. Liposomes entrapping $^3H-Ti plasmid$ were fused with Nicotiana sanderae protoplasts by treating with 5 mM $CaCl_2$ and 10% PEG. The fusion was evidenced by fluorescence microscopic technique. The amounts of Ti plasmid and PS associated with protoplasts were assayed by the radioactivity of $^3H-Ti plasmid$ and by the fluorescence of rhodamine B. About 7.9% of the PS liposome and 7.2% of PS-Chol liposome were fused with protoplasts. During the fusion process, about 30% of the liposomal contents of PS-Chol liposome was leaked, in contrast to about 60% leakage of its contents in PS liposome. Accounting the number of liposomes fused with protoplasts together with the encapsulation efficiency and the leakage of liposomal contents, it was calculated that ca. 1,700 Ti plasmid was transfered into one protoplast by the present method. This result may indicates that the present method transfers enough Ti plasmid into plant protoplast to elicit genetic transformation of plants.